Longer amplicons and greater yields with AccuTaq LA DNA polymerase

AccuTaq LA DNA polymerase (SA) and competitors' long and accurate enzymes were used to amplify 5 kb, 10 kb and 20 kb fragments of human genomic DNA.

Amplification conditions:
5 kb amplifications performed in 50 µl, 5 µl loaded onto the gel. For all samples 4 ng/µl of human genomic DNA was amplified with 0.05 units/µl of enzyme.
Cycling conditions: 94 °C 1 min, 30 cycles [94 °C 15 sec, 65 °C 20 sec, 72 °C 10 min] 72 °C 10 min final extension, hold at 4 °C
10 kb amplifications performed in 20 µl, 4 µl loaded onto the gel. For all samples 1 ng/µl of human genomic DNA was amplified.
Enzyme amounts:
AccuTaq LA: 0.05 units/µl
Supplier R: 0.075 units/µl
Supplier S: 0.1 units/µl
Cycling conditions: 94 °C 30 sec, 30 cycles [94 °C 5 sec, 62 °C 20 sec, 68 °C 20 min] 68 °C 20 min final extension, hold at 4 °C
15 kb amplifications performed in 20 µl, 4 µl loaded onto the gel. For all samples 1 ng/µl of human genomic DNA was amplified.
Enzyme amounts:
AccuTaq LA: 0.05 units/µl
Supplier R: 0.075 units/µl
Supplier S: 0.1 units/µl
Cycling conditions: 94 °C 30 sec, 30 cycles [94 °C 5 sec, 62 °C 20 sec, 68 °C 30 min] 68 °C 30 min final extension, hold at 4 °C

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